A549细胞三维立体培养模型的建立及耐药机制

The establishment of three-dimensional cell culture model of A549 and the drug tolerance mechanism

目的探讨肺腺癌A549细胞三维立体(3D)培养模型对顺铂(DDP)的耐药性及其黏附耐药机制。方法胶原培养条件下建立A549三维立体培养模型,MTT法检测平面(2D)培养和3D培养A549细胞敏感性。采用整合素β1激动剂(TSF)和抑制剂(MAB)干预,对其相关机制进行研究。结果 A549细胞3D培养模型中可见细胞生长良好,细胞之间黏附形成典型的多细胞球。2D培养模型中A549细胞对DDP的IC50值为12.84μg/ml;3D培养A549细胞对DDP的IC50是31.74μg/ml,为2D培养的2.47倍。与空白对照组比较,DDP干预组及MAB+DDP组的抑制率明显升高(P<0.01),TSF+DDP组有较显著差别(P<0.05);与DDP干预组相比较,TSF+DDP组细胞抑制率明显降低(P<0.05),MAB+DDP组明显升高(P<0.05)。结论胶原法成功建立A549细胞的3D培养模型。与2D培养相比,该细胞模型具有耐药的特性,其机制可能与促进整合素β1对细胞的黏附作用有关。

Objective To establish a three-dimensional culture model （3D）of human lung cancer A549 cells, evaluate the tolerance of DDP to the 3D ceils and study the cell adhesion mediated drug resistance（CAM-DR） mechanism. Methods The rat tail collagen solution was used to establish the 3D cell culture system, MTT method was used to detect the tolerance of 2D and 3D culture A549 cells to DDP, and discuss the CAM-DR mechanism. Results The improved method 3D culture system of A549 ceils was successfully established. The cells grew well and form some typical muhicellular spheres. The IC50 value of DDP on 2D cells culture model was 12.84 μg/ml;the IC50 value of DDP on 3D cells culture model was 31.74 μg/ml,2.47 times of the 2D cells culture model. Comparing with that of blank control group,the cell inhibition ratio of the DDP treatment group and the MAB＋DDP group were significantly higher（ P〈0.01 ）, and the TSF＋DDP group showed significant difference （P〈0.05）. Comparing with that of DDP treatment group, the cell inhibition ratio of the TSF＋DDP group was obviously lower（ P〈0.05 ）and the MAB＋DDP group significantly higher（ P〈0.05 ）. Conclusions The 3D A549 ceils culture model is established successfully using rat tail collagen solution. This model is more close to the tumor growing environment in vivo, and could reverse the adhesion-mediated drug resistance. The mechanism might be related with reducing the cells＇ adhesion mediated by the integrin β1.